The present invention relates to agents that act to antagonize the action of the glucagon peptide hormone. More particularly, it relates to glucagon antagonists or inverse agonists.
Glucagon is a key hormonal agent that, in co-operation with insulin, mediates homeostatic regulation of the amount of glucose in the blood. Glucagon primarily acts by stimulating certain cells (mostly liver cells) to release glucose when blood glucose levels fall. The action of glucagon is opposite to that of insulin, which stimulates cells to take up and store glucose whenever blood glucose levels rise. Both glucagon and insulin are peptide hormones.
Glucagon is produced in the alpha islet cells of the pancreas and insulin in the beta islet cells. Diabetes mellitus is a common disorder of glucose metabolism. The disease is characterized by hyperglycemia and may be classified as Type 1 diabetes, the insulin-dependent form, or Type 2 diabetes, which is non-insulin-dependent in character. Subjects with Type 1 diabetes are hyperglycemic and hypoinsulinemic, and the conventional treatment for this form of the disease is to provide insulin. However, in some patients with Type 1 or Type 2 diabetes, absolute or relative elevated glucagon levels have been shown to contribute to the hyperglycemic state. Both in healthy control animals as well as in animal models of Type 1 and Type 2 diabetes, removal of circulating glucagon with selective and specific antibodies has resulted in reduction of the glycemic level (Brand et al., Diabetologia 37, 985 (1994); Diabetes 43, [suppl 1], 172A (1994); Am. J. Physiol. 269, E469-E477 (1995); Diabetes 44 [suppl 1], 134A (1995); Diabetes 45, 1076 (1996)). These studies suggest that glucagon suppression or an action that antagonizes glucagon could be a useful adjunct to conventional antihyperglycemia treatment of diabetes. The action of glucagon can be suppressed by providing an antagonist or an inverse agonist, ie substances that inhibit or prevent glucagon induced responses. The antagonist can be peptidic or non-peptidic in nature. Native glucagon is a 29 amino acid peptide having the sequence:
His-Ser-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Ser-Arg-Arg-Ala-Gln-Asp-Phe-Val-Gln-Trp-Leu-Met-Asn-Thr-NH2.
Glucagon exerts its action by binding to and activating its receptor, which is part of the Glu-cagon-Secretin branch of the 7-transmembrane G-protein coupled receptor family (Jelinek et al., Science 259, 1614, (1993)). The receptor functions by an activation of the adenylyl cyclase second messenger system and the result is an increase in cAMP levels.
Several publications disclose peptides that are stated to act as glucagon antagonists. Probably, the most thoroughly characterized antagonist is DesHis1[Glu9]-glucagon amide (Unson et al., Peptides 10, 1171 (1989); Post et al., Proc. Natl. Acad. Sci. USA 90, 1662 (1993)). Other antagonists are eg DesHis1,Phe6[Glu9]-glucagon amide (Azizh et al., Bioorganic and Medicinal Chem. Lett. 16, 1849 (1995)) or NLeu9,Ala11,16-glucagon amide (Unson et al., J. Biol. Chem. 269(17), 12548 (1994)).
Peptide antagonists of peptide hormones are often quite potent; however, they are generally known not to be orally available because of degradation by physiological enzymes, and poor distribution in vivo. Therefore, orally available non-peptide antagonists of the peptide hormones are preferred. Among the non-peptide glucagon antagonists, a quinoxaline derivative, (2-styryl-3-[3-(dimethylamino)propylmethylamino]-6,7-dichloroquinoxaline was found to desplace glucagon from the rat liver receptor (Collins, J. L. et al., Bioorganic and Medicinal Chemistry Letters 2(9):915-918 (1992)). West, R. R. et al., WO 94/14426 (1994) discloses use of skyrin, a natural product comprising a pair of linked 9,10-anthracenedione groups, and its synthetic analogues, as glucagon antagonists. Anderson, P. L., U.S. Pat. No. 4,359,474 discloses the glucagon antagonistic properties of 1-phenyl pyrazole derivatives. Barcza, S., U.S. Pat. No. 4,374,130, discloses substituted disilacyclohexanes as glucagon antagonists. WO 98/04528 (Bayer Corporation) discloses substituted pyridines and biphenyls as glucagon antagonists. WO 97/16442 and U.S. Pat. No 5,776,954 (Merck and Co., Inc.) disclose substituted pyridyl pyrroles as glucagon antagonists and WO 98/21957, WO 98/22108, WO 98/22109 and U.S. Pat. No. 5,880,139 (Merck and Co., Inc.) disclose 2,4-diaryl-5-pyridylimidazoles as glucagon antagonists. Furthermore, WO 97/16442, U.S. Pat. Nos. 5,837,719 and 5,776,954 (Merck and Co., Inc.) discloses 2,5-substituted aryl pyrroles as glucagon antagonists. WO 98/24780, WO 98/24782, WO 99/24404 and WO 99/32448 (Amgen Inc.) disclose substituted pyrimidinone and pyridone compounds and substituted pyrimidine compounds, respectively, which are stated to posses glucagon antagonistic activity. Madsen et al (J. Med. Chem. 1998 (41) 5151-7) discloses a series of 2-(benzimidazol-2-ylthio)-1-(3,4-dihydroxyphenyl)-1-ethanones as competitive human glucagon receptor antagonists. WO 99/01423 (Novo Nordisk A/S) discloses a series of acylhydrazones as glucagon antagonists/inverse agonists.
These known glucagon antagonists differ structurally from the present compounds.
Definitions
The following is a detailed definition of the terms used to describe the compounds of the invention:
xe2x80x9cHalogenxe2x80x9d designates an atom selected from the group consisting of F, Cl, Br or I.
The term xe2x80x9cC1-6-alkylxe2x80x9d in the present context designates a branched or straight hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl, isohexyl and the like.
The term xe2x80x9cC2-6-alkenylxe2x80x9d as used herein represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one double bond. Examples of such groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3-butadienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5-hexenyl and the like.
The term xe2x80x9cC2-6-alkynylxe2x80x9d as used herein represents a branched or straight hydrocarbon group having from 2 to 6 carbon atoms and at least one triple bond. Examples of such groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 5-hexynyl, 2,4-hexadiynyl and the like.
The term xe2x80x9cC1-6-alkoxyxe2x80x9d as used herein, alone or in combination, refers to the radical xe2x80x94Oxe2x80x94C1-6-alkyl where C1-6-alkyl is as defined above. Representative examples are methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, sec-butoxy, tert-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like.
The term xe2x80x9cC1-6-alkanoylxe2x80x9d as used herein denotes a group xe2x80x94C(O)H or xe2x80x94C(O)xe2x80x94C1-5-alkyl. Representative examples are formyl, acetyl, propionyl, butyryl, valeryl, hexanoyl and the like.
The term xe2x80x9cC3-8-cycloalkylxe2x80x9d as used herein represents a carbocyclic group having from 3 to 8 carbon atoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
The term xe2x80x9cC3-8-cycloalkenylxe2x80x9d as used herein represents a carbocyclic group having from 3 to 8 carbon atoms containing a least one double bond. Representative examples are 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3-cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1,4-cyclooctadienyl and the like.
The term xe2x80x9cC4-8-cycloalkenylxe2x80x9d as used herein represents a carbocyclic group having from 4 to 8 carbon atoms containing a least one double bond. Representative examples are 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cyclohexenyl, 2-cyclohexenyl, 3-cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1,4-cyclooctadienyl and the like.
The term xe2x80x9cheterocyclylxe2x80x9d as used herein represents a saturated or partially unsaturated 3 to 10 membered ring containing one or more heteroatoms selected from nitrogen, oxygen and sulfur. Representative examples are pyrrolidinyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like.
The term xe2x80x9carylxe2x80x9d as used herein is intended to include carbocyclic aromatic ring systems such as phenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl, fluorenyl, indenyl, pentalenyl, azulenyl, and the like. Aryl is also intended to include the partially hydrogenated derivatives of the carbocyclic systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl and the like.
The term xe2x80x9caryloxyxe2x80x9d as used herein denotes a group xe2x80x94O-aryl, wherein aryl is as defined above.
The term xe2x80x9caroylxe2x80x9d as used herein denotes a group xe2x80x94C(O)-aryl, wherein aryl is as defined above.
The term xe2x80x9cheteroarylxe2x80x9d as used herein is intended to include heterocyclic aromatic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sulfur such as furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzothiophenyl (thianaphthenyl), indazolyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl and the like. Heteroaryl is also intended to include the partially hydrogenated derivatives of the heterocyclic systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 2,3-dihydrobenzofuranyl, pyrrolinyl, pyrazolinyl, indolinyl, oxazolidinyl, oxazolinyl, oxazepinyl and the like.
xe2x80x9cAryl-C1-6-alkylxe2x80x9d, xe2x80x9cheteroaryl-C1-6-alkylxe2x80x9d, xe2x80x9caryl-C2-6-alkenylxe2x80x9d etc. mean C1-6-alkyl or C2-6-alkenyl as defined above, substituted by an aryl or heteroaryl as defined above, for example: 
The term xe2x80x9coptionally substitutedxe2x80x9d as used herein means that the groups in question are either unsubstituted or substituted with one or more of the substituents specified. When the groups in question are substituted with more than one substituent the substituents may be the same or different.
Certain of the above defined terms may occur more than once in the structural formulae, and upon such occurrence each term shall be defined independently of the other.
Furthermore, when using the terms xe2x80x9cindependently arexe2x80x9d and xe2x80x9cindependently selected fromxe2x80x9d it should be understood that the groups in question may be the same or different.
The present invention is based on the unexpected observation that the compounds of the general formula (I) disclosed below antagonize the action of glucagon.
Accordingly, the invention is concerned with compounds of the general formula (I): 
wherein
V is xe2x80x94C(O)OR2, xe2x80x94C(O)NR2R3, xe2x80x94C(O)NR2OR3, xe2x80x94S(O)2OR2, 
xe2x80x83wherein
R2 and R3 independently are hydrogen or C1-6-alkyl,
R4 is hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR5, xe2x80x94NR5R6 or C1-6-alkyl,
wherein R5 and R6 independently are hydrogen or C1-6-alkyl,
A is 
xe2x80x83wherein
b is 0 or 1,
n is 0, 1, 2 or 3,
R7 is hydrogen, C1-6-alkyl or C3-8-cycloalkyl-C1-6-alkyl,
R8 and R9 independently are hydrogen or C1-6-alkyl,
Y is xe2x80x94C(O)xe2x80x94, xe2x80x94S(O)2xe2x80x94, xe2x80x94Oxe2x80x94 or a valence bond,
Z is phenylene or a divalent radical derived from a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur,
which may optionally be substituted with one or two groups R46 and R47 selected from hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR10, xe2x80x94NR10R11 and C1-6-alkyl,
wherein R10 and R11 independently are hydrogen or C1-6-alkyl,
or xe2x80x94Axe2x80x94Yxe2x80x94Zxe2x80x94 together are 
R1 is hydrogen or C1-6-alkyl,
X is 
xe2x80x83wherein
r is 0 or 1,
q and s independently are 0, 1, 2 or 3,
R12, R13, R14 and R15 independently are hydrogen or C1-6-alkyl,
D is 
xe2x80x83wherein
W is xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94S(O)2xe2x80x94, or xe2x80x94NR20xe2x80x94,
Wxe2x80x2 is xe2x95x90CR20xe2x80x2xe2x80x94 or xe2x95x90Nxe2x80x94,
R16, R17, R18 and R19 independently are
hydrogen, halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94NR21S(O)2R22, xe2x80x94S(O)2NR21R22, xe2x80x94S(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94OS(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94CH2C(O)NR21R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94CH2OR21, xe2x80x94CH2NR21R22, xe2x80x94OC(O)R21, xe2x80x94C(O)R21 or xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl or C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C3-8-cycloalkyl, C4-8-cycloalkenyl, heterocyclyl, C3-8-cycloalkyl-C1-6-alkyl, C3-8-cycloalkyl-C1-6-alkoxy, C3-8-cycloalkyloxy, C3-8-cycloalkyl-C1-6-alkylthio, C3-8-cycloalkylthio, C3-8-cycloalkyl-C2-6-alkenyl, C3-8-cycloalkyl-C2-6-alkynyl, C4-8-cycloalkenyl-C1-6-alkyl, C4-8-cycloalkenyl-C2-6-alkenyl, C4-8-cycloalkenyl-C2-6-alkynyl, heterocyclyl-C1-6-alkyl, heterocyclyl-C2-6-alkenyl or heterocyclyl-C2-6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from
xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkyl, aryl-C2-6-alkenyl, aryl-C2-6-alkynyl, heteroaryl, heteroaryl-C1-6-alkyl, heteroaryl-C2-6-alkenyl or heteroaryl-C2-6-alkynyl,
of which the aryl and heteroaryl moieties optionally may be substituted with one or more substituents selected from
halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94NR21S(O)2R22, xe2x80x94S(O)2NR21R22, xe2x80x94S(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94OS(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94CH2C(O)NR21R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94CH2OR21, xe2x80x94CH2NR21R22, xe2x80x94OC(O)R21, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
wherein R21 and R22 independently are hydrogen, xe2x80x94CF3, C1-6-alkyl, tri-C1-6-alkylsilyl, C3-8-cycloalkyl, C3-8-cycloalkyl-C1-6-alkyl, aryl, aryl-C1-6-alkyl or heteroaryl,
or R21 and R22 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
or two of the groups R16 to R19 when placed in adjacent positions together may form a bridge xe2x80x94(CR16xe2x80x2R17xe2x80x2)axe2x80x94Oxe2x80x94(CR18xe2x80x2R19xe2x80x2)cxe2x80x94Oxe2x80x94,
wherein
a is 0, 1 or 2,
c is 1 or 2,
R16xe2x80x2, R17xe2x80x2, R18xe2x80x2 and R19xe2x80x2 independently are hydrogen, C1-6-alkyl or halogen,
R20 and R20xe2x80x2 independently are hydrogen, C1-6-alkyl, C3-8-cycloalkyl or C3-8-cycloalkyl-C1-6-alkyl,
E is a 3 to 9 membered mono- or bicyclic ring which may optionally contain one or two double bonds and which may optionally contain one or two heteroatoms selected from nitrogen, oxygen and sulfur, wherein one or two groups R23 and R24 may be attached to the same or different ring carbon atoms and wherein a group R31 may be attached to a ring nitrogen atom when present, or 
xe2x80x83wherein
m and p independently are 0, 1, 2, 3 or 4, with the proviso that when both m and p are present in the same formula at least one of m and p is different from 0,
R23 and R24 independently are
hydrogen, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 or xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl or C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C3-8-cycloalkyl, C3-8-cycloalkylidene, C4-8-cycloalkenyl, heterocyclyl, C3-8-cycloalkyl-C1-6-alkyl, C3-8-cycloalkyl-C2-6-alkenyl, C3-8-cycloalkyl-C2-6-alkynyl, C4-8-cycloalkenyl-C1-6-alkyl, C4-8-cycloalkenyl-C2-6-alkenyl, C4-8-cycloalkenyl-C2-6-alkynyl, heterocyclyl-C1-6-alkyl, heterocyclyl-C2-6-alkenyl or heterocyclyl-C2-6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from
xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
aryl, aryloxy, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkyl, aryl-C2-6-alkenyl, aryl-C2-6-alkynyl, heteroaryl, heteroaryl-C1-6-alkyl, heteroaryl-C2-6-alkenyl or heteroaryl-C2-6-alkynyl,
of which the aryl and heteroaryl moieties optionally may be substituted with one or more substituents selected from
halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94NR36S(O)2R37, xe2x80x94S(O)2NR36R37, xe2x80x94S(O)NR36R37, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94OS(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94CH2C(O)NR36R37, xe2x80x94CH2C(O)NR36R37, xe2x80x94CH2OR36, xe2x80x94CH2NR36R37, xe2x80x94OC(O)R36, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
wherein R36 and R37 independently are hydrogen, C1-6-alkyl or aryl,
of which the aryl moiety optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR38, xe2x80x94NR38R39 and C1-6-alkyl,
wherein R38 and R39 independently are hydrogen or C1-6-alkyl,
or R36 and R37 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
or R23 and R24 when attached to the same ring carbon atom or different ring carbon atoms together may form a radical xe2x80x94Oxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Oxe2x80x94, xe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94 or xe2x80x94Sxe2x80x94(CH2)tCR40R41xe2x80x94(CH2)lxe2x80x94Sxe2x80x94,
wherein
t and l independently are 0, 1, 2, 3, 4 or 5,
R40 and R41 independently are hydrogen or C1-6-alkyl,
R25 to R30 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94NO2, xe2x80x94OR42, xe2x80x94NR42R43, C1-6-alkyl, C3-8-cycloalkyl or C4-8-cycloalkenyl,
wherein R42 and R43 independently are hydrogen or C1-6-alkyl, or
R42 and R43 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
R31, R32 and R33 independently are hydrogen or C1-6-alkyl,
R34 and R35 independently are
hydrogen, C1-6-alkyl, C1-6-alkoxy, C1-6-alkanoyl, xe2x80x94C(O)NR44R45 or xe2x80x94S(O)2R45,
aryl, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkanoyl or aryl-C1-6-alkyl,
of which the aryl moieties optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR44, xe2x80x94NR44R45 and C1-6-alkyl,
wherein R44 and R45 independently are hydrogen or C1-6-alkyl, or
R34 and R35 when attached to a carbon atom together with the said carbon atom may form a 3 to 8 membered cyclic ring optionally containing one or two heteroatoms selected from nitrogen, oxygen or sulfur, and optionally containing one or two double bonds, or
R34 and R35 when attached to a nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen or sulfur, and optionally containing one or two double bonds,
as well as any optical or geometric isomer or tautomeric form thereof including mixtures of these or a pharmaceutically acceptable salt thereof.
The compounds according to the invention preferably have an IC50 value of no greater than 5 xcexcM as determined by the Glucagon Binding Assay (I), Glucagon Binding Assay (II) or Glucagon Binding Assay (III) disclosed herein.
More preferably, the compounds according to the invention have a glucagon antagonistic activity as determined by the Glucagon Binding Assay (I), Glucagon Binding Assay (II) or Glucagon Binding Assay (III) disclosed herein corresponding to an IC50 value of less than 1 xcexcM, preferably of less than 500 nM and even more preferred of less than 100 nM.
The compounds according to the invention are useful for the treatment and/or prevention of an indication selected from the group consisting of hyperglycemia, IGT (impaired glucose tolerance), Type 2 diabetes, Type 1 diabetes and obesity.
In a further aspect the invention relates to compounds of the general formula (Ixe2x80x2): 
wherein
V is xe2x80x94C(O)OR2, xe2x80x94C(O)NR2R3, xe2x80x94C(O)NR2OR3, 
xe2x80x83wherein
R2 and R3 independently are hydrogen or C1-6-alkyl,
R4 is hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR5, xe2x80x94NR5R6 or C1-6-alkyl,
wherein R5 and R6 independently are hydrogen or C1-6-alkyl;
A is 
xe2x80x83wherein
b is 0 or 1,
n is 0, 1, 2 or 3,
R7 is hydrogen, C1-6-alkyl or C3-8-cycloalkyl-C1-6-alkyl,
R8 and R9 independently are hydrogen or C1-6-alkyl;
Y is xe2x80x94C(O)xe2x80x94, xe2x80x94S(O)2xe2x80x94 or xe2x80x94Oxe2x80x94;
Z is phenylene or a divalent radical derived from a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur,
which may optionally be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR10, xe2x80x94NR10R11 and C1-6-alkyl,
wherein R10 and R11 independently are hydrogen or C1-6-alkyl;
or xe2x80x94Axe2x80x94Yxe2x80x94Zxe2x80x94 together is 
R1 is hydrogen or C1-6-alkyl;
X is 
xe2x80x83wherein
r is 0 or 1,
q and s independently are 0, 1, 2 or 3,
R12, R13, R14 and R15 independently are hydrogen or C1-6-alkyl;
D is 
xe2x80x83wherein
W is xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 or xe2x80x94NR20xe2x80x94,
wherein R20 is hydrogen or C1-6-alkyl,
R16, R17, R18 and R19 independently are
hydrogen, halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94NR21S(O)2R22, xe2x80x94S(O)2NR21R22, xe2x80x94S(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94OS(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94CH2C(O)NR21R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94CH2OR21, xe2x80x94CH2NR21R22, xe2x80x94OC(O)R21, xe2x80x94C(O)R21 or xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl or C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C3-8-cycloalkyl, C3-8-cycloalkenyl, heterocyclyl, C3-8-cycloalkyl-C1-6-alkyl, C3-8-cycloalkyl-C2-6-alkenyl, C3-8-cycloalkyl-C2-6-alkynyl, C3-8-cycloalkenyl-C1-6-alkyl, C3-8-cycloalkenyl-C2-6-alkenyl, C3-8cycloalkenyl-C2-6-alkynyl, heterocyclyl-C1-6-alkyl, heterocyclyl-C2-6-alkenyl or heterocyclyl-C2-6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
aryl, aryloxy, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkyl, aryl-C2-6-alkenyl, aryl-C2-6-alkynyl, heteroaryl, heteroaryl-C1-6-alkyl, heteroaryl-C2-6-alkenyl or heteroaryl-C2-6-alkynyl,
of which the aryl and heteroaryl moieties optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94NR21S(O)2R22, xe2x80x94S(O)2NR21R22, xe2x80x94S(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94OS(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94CH2C(O)NR21R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94CH2OR21, xe2x80x94CH2NR21R22, xe2x80x94OC(O)R21, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
wherein R21 and R22 independently are hydrogen or C1-6-alkyl, or R21 and R22 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
or two of the groups R16 to R19 when placed in adjacent positions together may form a bridge xe2x80x94OCH2Oxe2x80x94 or xe2x80x94OCH2CH2Oxe2x80x94;
E is a 3 to 9 membered mono- or bicyclic ring which may optionally contain one or two double bonds and which may optionally contain one or two heteroatoms selected from nitrogen, oxygen and sulfur, wherein one or two groups R23 and R24 may be attached to the same or different ring carbon atoms and wherein a group R31 may be attached to a ring nitrogen atom when present, or 
xe2x80x83wherein
m and p independently ar 0, 1, 2, 3 or 4,
R23 and R24 independently are hydrogen, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 or xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6alkenyl or C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C3-8-cycloalkyl, C3-8-cycloalkenyl, heterocyclyl, C3-8-cycloalkyl-C1-6-alkyl, C3-8-cycloalkyl-C2-6-alkenyl, C3-8-cycloalkyl-C2-6-alkynyl, C3-8-cycloalkenyl-C1-6-alkyl, C3-8-cycloalkenyl-C2-6-alkenyl, C3-8-cycloalkenyl-C2-6-alkynyl, heterocyclyl-C1-6-alkyl, hetercyclyl-C2-6-alkeny or heterocyclyl-C2-6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
aryl, aryloxy, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkyl, aryl-C2-6-alkenyl, aryl-C2-6-alkynyl, heteroaryl, heteroaryl-C1-6-alkyl, heteroaryl-C2-6-alkenyl or heteroaryl-C2-6-alkynyl,
of which the aryl and heteroaryl moieties optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94NR36S(O)2R37, xe2x80x94S(O)2NR36R37, xe2x80x94S(O)NR36R37, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94OS(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94CH2C(O)NR36 R37, xe2x80x94CH2C(O)NR36R37, xe2x80x94CH2OR36, xe2x80x94CH2NR36R37, xe2x80x94OC(O)R36, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
wherein R36 and R37 independently are hydrogen, C1-6-alkyl or aryl,
of which the aryl moiety optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR38, xe2x80x94NR38R39 and C1-6-alkyl,
wherein R38 and R39 independently are hydrogen or C1-6-alkyl, or R36 and R37 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
or R23 and R24 when attached to the same ring carbon atom or different ring carbon atoms together may form a radical xe2x80x94Oxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Oxe2x80x94, xe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94 or xe2x80x94Sxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Sxe2x80x94,
wherein
t and l independently are 0, 1, 2, 3, 4 or 5,
R40 and R41 independently are hydrogen or C1-6-alkyl,
R25 to R30 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94NO2, xe2x80x94OR42, xe2x80x94NR42R43, C1-6-alkyl or C3-8-cycloalkyl,
wherein R42 and R43 independently are hydrogen or C1-6-alkyl,
R31, R32 and R33 independently are hydrogen or C1-6-alkyl,
R34 and R35 independently are hydrogen, C1-6-alkyl, C1-6-alkoxy, C1-6-alkanoyl, xe2x80x94C(O)NR44R45 or xe2x80x94S(O)2R45,
aryl, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkanoyl or aryl-C1-6-alkyl,
of which the aryl moieties optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR44, xe2x80x94NR44R45 and C1-6-alkyl,
wherein R44 and R45 independently are hydrogen or C1-6-alkyl,
or R34 and R35 when attached to the carbon atom together with the said carbon atom may form a 3 to 8 membered cyclic ring optionally containing one or two heteroatoms selected from nitrogen, oxygen or sulfur, and optionally containing one or two double bonds,
or R34 and R35 when attached to the nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen or sulfur, and optionally containing one or two double bonds;
as well as any optical or geometric isomer or tautomeric form thereof including mixtures of these or a pharmaceutically acceptable salt thereof;
having an IC50 value of no greater than 5 xcexcM as determined by the Glucagon Binding Assay (I), Glucagon Binding Assay (II) or Glucagon Binding Assay (III) disclosed herein.
By a compound of the general formula (Ixe2x80x2) having an IC50 value of no greater than 5 xcexcM as determined by the Glucagon Binding Assay (I), Glucagon Binding Assay (II) or Glucagon Binding Assay (III) is meant any compound of the defined formula having such activity, without limitation to any utility or usefulness for treating any specific indication.
In a further aspect the invention relates to compounds of the general formula (Ixe2x80x3): 
wherein
V is xe2x80x94C(O)OR2, xe2x80x94C(O)NR2R3, xe2x80x94C(O)NR2OR3, xe2x80x94S(O)2OR2, 
xe2x80x83wherein
R2 and R3 independently are hydrogen or C1-6-alkyl,
R4 is hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR5, xe2x80x94NR5R6 or C1-6-alkyl,
wherein R5 and R6 independently are hydrogen or C1-6-alkyl,
A is 
xe2x80x83wherein
b is 0 or 1,
n is 0, 1, 2 or 3,
R7 is hydrogen, C1-6-alkyl or C3-8-cycloalkyl-C1-6-alkyl,
R8 and R9 independently are hydrogen or C1-6-alkyl,
Y is xe2x80x94C(O)xe2x80x94, xe2x80x94S(O)2xe2x80x94, xe2x80x94Oxe2x80x94 or a valence bond,
Z is phenylene or a divalent radical derived from a 5 or 6 membered heteroaromatic ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur,
which may optionally be attached to one or two groups R46 and R47 selected from hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR10, xe2x80x94NR10R11 and C1-6-alkyl,
wherein R10 and R11 independently are hydrogen or C1-6-alkyl, or xe2x80x94Axe2x80x94Yxe2x80x94Zxe2x80x94 together are 
R1 is hydrogen or C1-6-alkyl,
X is 
xe2x80x83wherein
r is 0 or 1,
q and s independently are 0, 1, 2 or 3,
R12, R13, R14 and R15 independently are hydrogen or C1-6-alkyl,
D is 
xe2x80x83wherein
W is xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94, xe2x80x94S(O)2xe2x80x94 or xe2x80x94NR20xe2x80x94,
Wxe2x80x2 is xe2x95x90CR20xe2x80x2xe2x80x94 or xe2x95x90Nxe2x80x94,
R20 and R20xe2x80x2 are hydrogen, C1-6-alkyl or C3-8-cycloalkyl-C1-6-alkyl,
R16, R17, R18 and R19 independently are
hydrogen, halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94NR21S(O)2R22, xe2x80x94S(O)2NR21R22, xe2x80x94S(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94OS(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94CH2C(O)NR21R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94CH2OR21, xe2x80x94CH2NR21R22, xe2x80x94OC(O)R21, xe2x80x94C(O)R21 or xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl or C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C3-8-cycloalkyl, C3-8-cycloalkenyl, heterocyclyl, C3-8-cycloalkyl-C1-6-alkyl, C3-8-cycloalkyl-C1-6-alkoxy, C3-8-cycloalkyloxy, C3-8-cycloalkyl-C1-6-alkylthio, C3-8-cycloalkylthio, C3-8-cycloalkyl-C2-6-alkenyl, C3-8-cycloalkyl-C2-6-alkynyl, C3-8-cycloalkenyl-C1-6-alkyl, C3-8-cycloalkenyl-C2-6-alkenyl, C3-8-cycloalkenyl-C2-6-alkynyl, heterocyclyl-C1-6-alkyl, heterocyclyl-C2-6-alkenyl or heterocyclyl-C2-6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from
xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkyl, aryl-C2-6-alkenyl, aryl-C2-6-alkynyl, heteroaryl, heteroaryl-C1-6-alkyl, heteroaryl-C2-6-alkenyl or heteroaryl-C2-6-alkynyl,
of which the aryl and heteroaryl moieties optionally may be substituted with one or more substituents selected from
halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94NR21S(O)2R22, xe2x80x94S(O)2NR21R22, xe2x80x94S(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94OS(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94CH2C(O)NR21R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94CH2OR21, xe2x80x94CH2NR21R22, xe2x80x94OC(O)R21, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR21, xe2x80x94NR21R22, xe2x80x94SR21, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94C(O)NR21R22, xe2x80x94OC(O)NR21R22, xe2x80x94NR21C(O)R22, xe2x80x94OCH2C(O)NR21R22, xe2x80x94C(O)R21 and xe2x80x94C(O)OR21,
wherein R21 and R22 independently are hydrogen, xe2x80x94CF3, C1-6-alkyl, tri-C1-6-alkylsilyl, C3-8-cycloalkyl, C3-8-cycloalkyl-C1-6-alkyl, aryl, aryl-C1-6-alkyl or heteroaryl, or R21 and R22 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
or two of the groups R16 to R19 when placed in adjacent positions together may form a bridge xe2x80x94(CR16xe2x80x2R17xe2x80x2)axe2x80x94Oxe2x80x94(CR18xe2x80x2R19xe2x80x2)cxe2x80x94Oxe2x80x94,
wherein
a is 0, 1 or 2,
c is 1 or 2,
R16xe2x80x2, R17xe2x80x2, R18xe2x80x2 and R19xe2x80x2 independently are hydrogen, C1-6-alkyl or halogen,
E is a 3 to 9 membered mono- or bicyclic ring which may optionally contain one or two double bonds and which may optionally contain one or two heteroatoms selected from nitrogen, oxygen and sulfur, wherein one or two groups R23 and R24 may be attached to the same or different ring carbon atoms and wherein a group R31 may be attached to a ring nitrogen atom when present, or 
xe2x80x83wherein
m and p independently are 0, 1, 2, 3 or 4, with the proviso that when both m and p are present in the same formula at least one of m and p is different from 0,
R23 and R24 independently are
hydrogen, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 or xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl or C2-6-alkynyl,
which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C3-8-cycloalkyl, C3-8-cycloalkylidene, C3-8-cycloalkenyl, heterocyclyl, C3-8-cycloalkyl-C1-6-alkyl, C3-8-cycloalkyl-C2-6-alkenyl, C3-8-cycloalkyl-C2-6-alkynyl, C3-8-cycloalkenyl-C1-6-alkyl, C3-8-cycloalkenyl-C2-6-alkenyl, C3-8-cycloalkenyl-C2-6-alkynyl, heterocyclyl-C1-6-alkyl, heterocyclyl-C2-6-alkenyl or heterocyclyl-C2-6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substituents selected from
xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
aryl, aryloxy, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkyl, aryl-C2-6-alkenyl, aryl-C2-6-alkynyl, heteroaryl, heteroaryl-C1-6-alkyl, heteroaryl-C2-6-alkenyl or heteroaryl-C2-6-alkynyl,
of which the aryl and heteroaryl moieties optionally may be substituted with one or more substituents selected from
halogen, xe2x80x94CN, xe2x80x94CH2CN, xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94OS(O)2CF3, xe2x80x94SCF3, xe2x80x94NO2, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94NR36S(O)2R37, xe2x80x94S(O)2NR36R37, xe2x80x94S(O)NR36R37, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94OS(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94CH2C(O)NR36R37, xe2x80x94CH2C(O)NR36R37, xe2x80x94CH2OR36, xe2x80x94CH2NR36R37, xe2x80x94OC(O)R36, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
C1-6-alkyl, C2-6-alkenyl and C2-6-alkynyl,
xe2x80x83which may optionally be substituted with one or more substituents selected from xe2x80x94CHF2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OCHF2, xe2x80x94OCH2CF3, xe2x80x94OCF2CHF2, xe2x80x94SCF3, xe2x80x94OR36, xe2x80x94NR36R37, xe2x80x94SR36, xe2x80x94S(O)R36, xe2x80x94S(O)2R36, xe2x80x94C(O)NR36R37, xe2x80x94OC(O)NR36R37, xe2x80x94NR36C(O)R37, xe2x80x94OCH2C(O)NR36R37, xe2x80x94C(O)R36 and xe2x80x94C(O)OR36,
wherein R36 and R37 independently are hydrogen, C1-6-alkyl or aryl,
of which the aryl moiety optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94OR38, xe2x80x94NR33R39 and C1-6-alkyl,
wherein R38 and R39 independently are hydrogen or C1-6-alkyl,
or R36 and R37 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
or R23 and R24 when attached to the same ring carbon atom or different ring carbon atoms together may form a radical xe2x80x94Oxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Oxe2x80x94, xe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94 or xe2x80x94Sxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Sxe2x80x94,
wherein
t and l independently are 0, 1, 2, 3, 4 or 5,
R40 and R41 independently are hydrogen or C1-6-alkyl,
R25 to R30 independently are hydrogen, halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94NO2, xe2x80x94OR42, xe2x80x94NR42R43, C1-6-alkyl, C3-8-cycloalkyl or C3-8-cycloalkenyl,
wherein R42 and R43 independently are hydrogen or C1-6-alkyl, or
R42 and R43 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulfur, and optionally containing one or two double bonds,
R31, R32 and R33 independently are hydrogen or C1-6-alkyl,
R34 and R35 independently are
hydrogen, C1-6-alkyl, C1-6-alkoxy, C1-6-alkanoyl, xe2x80x94C(O)NR44R45 or xe2x80x94S(O)2R45,
aryl, aroyl, aryl-C1-6-alkoxy, aryl-C1-6-alkanoyl or aryl-C1-6-alkyl,
of which the aryl moieties optionally may be substituted with one or more substituents selected from halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94OR44, xe2x80x94NR44R45 and C1-6-alkyl,
wherein R44 and R45 independently are hydrogen or C1-6-alkyl, or
R34 and R35 when attached to the carbon atom together with the said carbon atom may form a 3 to 8 membered cyclic ring optionally containing one or two heteroatoms selected from nitrogen, oxygen or sulfur, and optionally containing one or two double bonds, or
R34 and R35 when attached to the nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen or sulfur, and optionally containing one or two double bonds,
as well as any optical or geometric isomer or tautomeric form thereof including mixtures of these or a pharmaceutically acceptable salt thereof.
In a preferred embodiment V is xe2x80x94C(O)OH, xe2x80x94S(O)2OH, xe2x80x94C(O)NHOH or 5-tetrazolyl, and more preferred xe2x80x94C(O)OH or 5-tetrazolyl.
In a preferred embodiment A is 
wherein R7 is as defined for formula (I).
More preferred A is 
In a preferred embodiment Y is xe2x80x94C(O)xe2x80x94.
In another preferred embodiment Y is a valence bond.
In a preferred embodiment Z is 
wherein R46 and R47 are as defined for formula (I).
More preferably, Z is 
In a preferred embodiment R1 is hydrogen.
In another preferred embodiment R1 is methyl.
In a preferred embodiment X is 
wherein q, r, s, R12, R13 and R14 are as defined for formula (I).
More preferably, X is 
wherein q is 0 or 1, r is 0 or 1, s is 0, 1 or 2, and R13 is hydrogen or C1-6-alkyl.
Even more preferably, X is xe2x80x94C(O)NHxe2x80x94, xe2x80x94C(O)NHCH2xe2x80x94, xe2x80x94C(O)NHCH(CH3)xe2x80x94, xe2x80x94C(O)NHCH2CH2xe2x80x94, xe2x80x94C(O)CH2xe2x80x94, xe2x80x94C(O)CHxe2x95x90CHxe2x80x94, xe2x80x94(CH2)sxe2x80x94, xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)Oxe2x80x94 or xe2x80x94NHC(O)xe2x80x94, wherein s is 0 or 1.
Still more preferably, X is xe2x80x94C(O)NHxe2x80x94, xe2x80x94C(O)NHCH2xe2x80x94, xe2x80x94C(O)NHCH(CH3)xe2x80x94, xe2x80x94C(O)NHCH2CH2xe2x80x94, xe2x80x94C(O)CH2xe2x80x94, xe2x80x94(CH2)xe2x80x94, xe2x80x94C(O)xe2x80x94 or xe2x80x94NHC(O)xe2x80x94.
Of these X is preferably xe2x80x94C(O)NHxe2x80x94 or xe2x80x94C(O)NHCH(CH3)xe2x80x94.
In a preferred embodiment D is 
wherein R16, R17, R18, R19 and R20 are as defined for formula (I).
More preferably, D is 
wherein R16, R17, R18 and R20 are as defined for formula (I).
In a preferred embodiment D is 
wherein R16, R17 and R20 are as defined for formula (I).
More preferably, R16 and R17 are both hydrogen and R20 is C1-6-alkyl or C3-8-cycloalkyl-C1-6-alkyl. Even more preferably, R20 is cyclopropylmethyl, butyl or isopropyl, especially preferred isopropyl.
In another preferred embodiment D is 
wherein R16 and R17 are as defined for formula (I).
In yet another preferred embodiment D is 
wherein R16, R17 and R18 are as defined for formula (I).
Preferably, R16, R17 and R18 are independently
hydrogen, halogen, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CF3, xe2x80x94OCF3, hydroxy, xe2x80x94SCF3, C1-6-alkyl, C1-6-alkyl substituted with hydroxy, C1-6-alkyl substituted with xe2x80x94S(O)2R21, C1-6-alkoxy, xe2x80x94Sxe2x80x94C1-6-alkyl, xe2x80x94C(O)OR21, xe2x80x94C(O)R21, xe2x80x94CH2(O)R21, xe2x80x94C(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94S(O)2CF3, xe2x80x94S(O)2NR21R22, C3-8-cycloalkyl, C3-8-cycloalkyl-C1-6-alkoxy, C3-8-cycloalkyl-C1-6-alkylthio or C3-8-cycloalkylthio
wherein R21 and R22 independently are hydrogen, C1-6-alkyl, tri-C1-6-alkylsilyl, C3-8-cycloalkyl, C3-8-cycloalkyl-C1-6-alkyl, phenyl, phenyl-C1-6-alkyl, 2,3-dihydroindolyl or isoindolyl, or R21 and R22 together with the nitrogen atom to which they are attached form a piperidine ring,
phenoxy, phenoxycarbonyl, phenyl, phenyl-C1-6-alkoxy, phenyl-C1-6-alkyl, furanyl, tetrazolyl, benzoxazolyl or oxadiazolyl, of which the ring systems optionally may be substituted with halogen, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94NO2, xe2x80x94C(O)OR21, xe2x80x94OR21, xe2x80x94NR21R22 or C1-6-alkyl, wherein R21 and R22 independently are hydrogen or C1-6-alkyl, or
wherein R16 and R17 in adjacent positions form the radical xe2x80x94Oxe2x80x94CH2xe2x80x94Oxe2x80x94, xe2x80x94CF2xe2x80x94Oxe2x80x94CF2xe2x80x94Oxe2x80x94 or xe2x80x94Oxe2x80x94CF2xe2x80x94CF2xe2x80x94Oxe2x80x94, and R18 is hydrogen.
More preferably, R16, R17 and R18 are independently
hydrogen, halogen, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94SCF3, C1-6-alkyl, C1-6-alkyl substituted with hydroxy, C1-6-alkyl substituted with xe2x80x94S(O)2R21, C1-6-alkoxy, xe2x80x94Sxe2x80x94C1-6-alkyl, xe2x80x94C(O)OR21, xe2x80x94C(O)R21, xe2x80x94CH2(O)R21, xe2x80x94C(O)NR21R22, xe2x80x94S(O)R21, xe2x80x94S(O)2R21, xe2x80x94S(O)2CF3, xe2x80x94S(O)2NR21R22, C3-8-cycloalkyl-C1-6-alkoxy, C3-8-cycloalkyl-C1-6-alkylthio or C3-8-cycloalkylthio,
wherein R21 and R22 independently are hydrogen, C1-6-alkyl, tri-C1-6alkylsilyl, C3-8-cycloalkyl, C3-8-cycloalkyl-C1-6-alkyl, phenyl or 2,3-dihydroindolyl, or R21 and R22 together with the nitrogen atom to which they are attached form a piperidine ring,
phenoxy, phenyl, benzyl, furanyl, tetrazolyl, benzoxazolyl or oxadiazolyl, of which the ring systems optionally may be substituted with halogen, xe2x80x94C(O)OR21 or C1-6-alkyl, wherein R21 is hydrogen or C1-6-alkyl, or
wherein R16 and R17 in adjacent positions form the radical xe2x80x94CF2xe2x80x94Oxe2x80x94CF2xe2x80x94Oxe2x80x94 or xe2x80x94Oxe2x80x94CF2xe2x80x94CF2xe2x80x94Oxe2x80x94, and R18 is hydrogen.
Even more preferably, R16, R17 and R18 independently are
hydrogen, halogen, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CF3, xe2x80x94OCF3, xe2x80x94SCF3, C1-6-alkyl, C1-6-alkyl substituted with hydroxy, C1-6alkoxy, xe2x80x94Sxe2x80x94C1-6-alkyl, xe2x80x94C(O)OR21, xe2x80x94C(O)R21, xe2x80x94CH2(O)R21, xe2x80x94C(O)NR21R22, xe2x80x94S(O)2R21, xe2x80x94(O)2CF3 or xe2x80x94S(O)2NR21R22,
wherein R21 and R22 independently are hydrogen, C1-6-alkyl, tri-C1-6-alkylsilyl, phenyl or 2,3-dihydroindolyl,
phenoxy, phenyl, benzyl, furanyl, tetrazolyl, benzoxazolyl or oxadiazolyl, of which the ring systems optionally may be substituted with halogen, xe2x80x94C(O)OR21 or C1-6-alkyl, wherein R21 is hydrogen or C1-6-alkyl, or
wherein R16 and R17 in adjacent positions form the radical xe2x80x94CF2xe2x80x94Oxe2x80x94CF2xe2x80x94Oxe2x80x94 or xe2x80x94Oxe2x80x94CF2xe2x80x94CF2xe2x80x94Oxe2x80x94, and R18 is hydrogen.
Still more preferably, R16, R17 and R18 are independently hydrogen, halogen, xe2x80x94CN, xe2x80x94NO2, xe2x80x94CFxe2x80x943, xe2x80x94OCF3, xe2x80x94SCF3, C1-6-alkyl, C1-6-alkoxy, xe2x80x94Sxe2x80x94C1-6-alkyl, xe2x80x94C(O)OC1-6-alkyl, xe2x80x94S(O)2C1-6-alkyl, xe2x80x94S(O)2CF3, xe2x80x94C(O)N(C1-6alkyl)(C1-6-alkyl), xe2x80x94S(O)2N(phenyl)(C1-6-alkyl), xe2x80x94C(xe2x95x90O)C1-6-alkyl, xe2x80x94CH2OH, xe2x80x94CH2O(tri-C1-6-alkylsilyl), 2,3-dihydroindol-1-ylsulfonyl, phenoxy, phenyl, 4-chlorophenyl, 1,3,5-trimethylbenzyl, benzoxazolyl, 2-methyltetrazol-5-yl, 2-methyl-3-methoxycarbonylfuran-5-yl or 3-isopropyl-[1,2,4]oxadiazol-5-yl).
In a preferred embodiment one of R16 to R18 is hydrogen.
In another preferred embodiment two of R16 to R18 are hydrogen.
In yet another preferred embodiment R16 and R17 are both hydrogen and R18 is xe2x80x94OCF3, xe2x80x94SCF3xe2x80x94CF3, xe2x80x94S(O)2CH3, phenyl, halogen, C1-6-alkyl, nitro, xe2x80x94Sxe2x80x94C1-6-alkyl or xe2x80x94S(O)2NR21R22, wherein R21 is C1-6-alkyl and R22 is phenyl.
In still another preferred embodiment R16 and R17 are both hydrogen and R18 is xe2x80x94OCF3 or halogen.
In a further embodiment R16 is hydrogen and R17 and R18 are both halogen or are both xe2x80x94CF3.
In yet a further embodiment R16 is hydrogen, R17 is xe2x80x94CF3 and R18 is halogen, xe2x80x94CN, C1-6-alkoxy or xe2x80x94OCF3.
In still a further embodiment R16 is hydrogen, R17 is xe2x80x94OCF3 and R18 is xe2x80x94S(O)2CH3, xe2x80x94CH2O-tri-C1-6-alkylsilyl, benzoxazolyl or xe2x80x94CH2OH.
In another embodiment R16 is hydrogen, R17 is C1-6-alkyl and R18 is xe2x80x94S(O)2NR21R22, wherein R21 is C1-6-alkyl and R22 is phenyl.
In still another embodiment R16, R17 and R18 are selected from hydrogen, xe2x80x94OCF3, xe2x80x94CF3, xe2x80x94Br, xe2x80x94F and xe2x80x94Cl.
In a preferred embodiment E is 
wherein
m, p and R23 to R35 are as defined for formula (I).
Preferably, E is 
wherein m, p and R23 to R35 are as defined for formula (I).
More preferably, E is 
wherein p, R23, R24, R25, R26, R27, R28 , R29, R30, R34 and R35 are as defined for formula (I).
Even more preferably, E is 
wherein R23, R24, R25, R26, R27, R34 and R35 are as defined for formula (I).
When E is 
R34 and R35 are preferably independently C1-6-alkyl, hydrogen or C1-6-alkoxy. More preferably, R34 and R35 are both C1-6-alkyl.
In another preferred embodiment E is 
wherein R23 and R24 are as defined for formula (I).
Preferably, E is 
wherein R23 and R24 are as defined for formula (I).
Preferably, R23 and R24 are independently selected from hydrogen, C1-6-alkyl, C3-8-cycloalkyl, C3-8-cycloalkylidene, phenoxy, phenyl, xe2x80x94C(O)NR36R37 and xe2x80x94OC(O)NH-phenyl, of which the phenyl moiety optionally may be substituted with xe2x80x94OCF3, wherein R36 and R37 are as defined in claim 1, or R23 and R24 together form the radical xe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94, xe2x80x94Oxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94(CH2)txe2x80x94CR40R41xe2x80x94(CH2)lxe2x80x94Sxe2x80x94, wherein t, l, R40 and R41 are as defined for formula (I).
More preferably, R23 is hydrogen and R24 is C1-6-alkyl such as tert-butyl or C3-8-cycloalkyl such as cyclohexyl, wherein R23 and R24 are both C1-6-alkyl or wherein R23 and R24 together form the radical xe2x80x94(CH2)5xe2x80x94.
In yet another preferred embodiment E is 
wherein R25, R26 and R27 are as defined for formula (I).
Preferably, E is 
wherein R25, R26 and R27 are as defined for formula (I).
Preferably, R25, R26 and R27 are independently selected from hydrogen, halogen, C1-6-alkyl, C1-6-alkoxy, C3-8-cycloalkyl, C4-8-cycloalkenyl, xe2x80x94CF3, xe2x80x94OCF3 or xe2x80x94NR42R43, wherein R42 and R43 are as defined for formula (I).
More preferably, E is 
wherein R25 is xe2x80x94OCF3, xe2x80x94CF3, C1-6-alkyl such as tert-butyl, piperidyl, C3-8-cycloalkyl such as cyclohexyl or C4-8-cycloalkenyl such as cyclohexenyl.
In another preferred embodiment E is 
Of these E is preferably 
In one preferred embodiment the present invention relates to compounds of the general formula (I1): 
wherein V, A, R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In another preferred embodiment the present invention relates to compounds of the general formula (I2): 
wherein V, A, R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In still another preferred embodiment the present invention relates to compounds of the general formula (I3): 
wherein V, A, R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In yet another preferred embodiment the present invention relates to compounds of the general formula (I4): 
wherein V is xe2x80x94C(O)OR2, xe2x80x94C(O)NR2R3 or xe2x80x94C(O)NR2OR3, and R1, R2, R3, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In a further preferred embodiment the present invention relates to compounds of the general formula (I5): 
wherein R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In still a further preferred embodiment the present invention relates to compounds of the general formula (I6): 
wherein R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In yet a further preferred embodiment the present invention relates to compounds of the general formula (I7): 
wherein R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In another preferred embodiment the present invention relates to compounds of the general formula (I8): 
wherein R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In still another preferred embodiment the present invention relates to compounds of the general formula (I9): 
wherein R46, R47, R1, E, X and D are as defined for formula (I) or in any one of the above preferred embodiments.
In the above formulae (I1) to (I3) and (I5) to (I9), R46 and R47 are preferably both hydrogen.
The compounds of the present invention may have one or more asymmetric centres and it is intended that any optical isomers, as separated, pure or partially purified optical isomers or racemic mixtures thereof are included within the scope of the invention.
Furthermore, when a double bond or a fully or partially saturated ring system is present in the molecule geometric isomers may be formed. It is intended that any geometric isomers, as separated, pure or partially purified geometric isomers or mixtures thereof are included within the scope of the invention. Likewise, molecules having a bond with restricted rotation may form geometric isomers. These are also intended to be included within the scope of the present invention.
Furthermore, some of the compounds of the present invention may exist in different tautomeric forms and it is intended that any tautomeric forms which the compounds are able to form are included within the scope of the present invention.
The present invention also encompasses pharmaceutically acceptable salts of the present compounds. Such salts include pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts, ammonium and alkylated ammonium salts. Acid addition salts include salts of inorganic acids as well as organic acids. Representative examples of suitable inorganic acids include hydrochloric, hydrobromic, hydroiodic, phosphoric, sulfuric, nitric acids and the like. Representative examples of suitable organic acids include formic, acetic, trichloroacetic, trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric, glycolic, lactic, maleic, malic, malonic, mandelic, oxalic, picric, pyruvic, salicylic, succinic, methanesulfonic, ethanesulfonic, tartaric, ascorbic, pamoic, bismethylene salicylic, ethanedisulfonic, gluconic, citraconic, aspartic, stearic, palmitic, EDTA, glycolic, p-aminobenzoic, glutamic, benzenesulfonic, p-toluenesulfonic acids and the like. Further examples of pharmaceutically acceptable inorganic or organic acid addition salts include the pharmaceutically acceptable salts listed in J. Pharm. Sci. 1977, 66, 2, which is incorporated herein by reference. Examples of metal salts include lithium, sodium, potassium, magnesium salts and the like. Examples of ammonium and alkylated ammonium salts include ammonium, methyl-, dimethyl-, trimethyl-, ethyl-, hydroxyethyl-, diethyl-, butyl-, tetramethylammonium salts and the like.
Also intended as pharmaceutically acceptable acid addition salts are the hydrates, which the present compounds, are able to form.
Furthermore, the pharmaceutically acceptable salts comprise basic amino acid salts such as lysine, arginine and ornithine.
The acid addition salts may be obtained as the direct products of compound synthesis. In the alternative, the free base may be dissolved in a suitable solvent containing the appropriate acid, and the salt isolated by evaporating the solvent or otherwise separating the salt and solvent.
The compounds of the present invention may form solvates with standard low molecular weight solvents using methods well known to the person skilled in the art. Such solvates are also contemplated as being within the scope of the present invention.
The invention also encompasses prodrugs of the present compounds, which on administration undergo chemical conversion by metabolic processes before becoming pharmacologically active substances. In general, such prodrugs will be functional derivatives of the compounds of the general formula (I), which are readily convertible in vivo into the required compound of the formula (I). Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in xe2x80x9cDesign of Prodrugsxe2x80x9d, ed. H. Bundgaard, Elsevier, 1985.
The invention also encompasses active metabolites of the present compounds.
The compounds according to the present invention act to antagonize the action of glucagon and are accordingly useful for the treatment and/or prevention of disorders and diseases in which such an antagonism is beneficial.
Accordingly, in a further aspect the invention relates to a compound according to the invention for use as a medicament.
The invention also relates to pharmaceutical compositions comprising, as an active ingredient, at least one compound according to the invention together with one or more pharmaceutically acceptable carriers or excipients.
The pharmaceutical composition is preferably in unit dosage form, comprising from about 0.05 mg to about 1000 mg, preferably from about 0.1 mg to about 500 mg and especially preferred from about 0.5 mg to about 200 mg of the compound according to the invention.
Furthermore, the invention relates to the use of a compound according to the invention for the preparation of a pharmaceutical composition for the treatment and/or prevention of a disorder or disease, wherein a glucagon antagonistic action is beneficial.
The invention also relates to a method for the treatment and/or prevention of disorders or diseases, wherein a glucagon antagonistic action is beneficial the method comprising administering to a subject in need thereof an effective amount of a compound according to the invention.
Owing to their antagonizing effect of the glucagon receptor the present compounds may be suitable for the treatment and/or prevention of any glucagon-mediated conditions and diseases.
Accordingly, the present compounds may be applicable for the treatment and/or prevention of hyperglycemia, IGT, insulin resistance syndromes, syndrome X, Type 1 diabetes, Type 2 diabetes, hyperlipidemia, dyslipidemia, hypertriglyceridemia, glucagonomas, acute pancreatitis, cardiovascular diseases, hypertension, cardiac hypertrophy, gastrointestinal disorders, obesity, diabetes as a consequence of obesity, diabetic dyslipidemia, etc. Furthermore, they may be applicable as diagnostic agents for identifying patients having a defect in the glucagon receptor, as a therapy to increase gastric acid secretions and to reverse intestinal hypomobility due to glucagon administration.
In a preferred embodiment of the invention the present compounds are used for the manufacture of a medicament for the treatment and/or prevention of hyperglycemia.
In yet a preferred embodiment of the invention the present compounds are used for the manufacture of a medicament for lowering blood glucose in a mammal.
In another preferred embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the treatment and/or prevention of IGT.
In still another preferred embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the treatment and/or prevention of Type 2 diabetes.
In yet another preferred embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the delaying or prevention of the progression from IGT to Type 2 diabetes.
In yet another preferred embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the delaying or prevention of the progression from non-insulin requiring Type 2 diabetes to insulin requiring Type 2 diabetes.
In a further preferred embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the treatment and/or prevention of Type 1 diabetes. Such treatment and/or prevention is normally accompanied by insulin therapy.
In a further preferred embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the treatment and/or prevention of obesity.
In still a further embodiment of the invention the present compounds are used for the preparation of a pharmaceutical composition for the treatment and/or prevention of an appetite regulation or energy expenditure disorder.
In a further aspect of the invention the present compounds may be administered in combination with one or more pharmacologically active substances eg selected from antidiabetics, antiobesity agents, antihypertensive agents and agents for the treatment and/or prevention of complications resulting from or associated with diabetes.
Suitable antidiabetics comprise insulin, GLP-1 derivatives such as those disclosed in WO 98/08871 to Novo Nordisk ANS, which is incorporated herein by reference, as well as orally active hypoglycaemic agents.
The orally active hypoglycaemic agents preferably comprise sulphonylureas, biguanides, meglitinides, oxadiazolidinediones, thiazolidinediones, glucosidase inhibitors, glucagon antagonists, GLP-1 agonists, potassium channel openers such as those disclosed in WO 97/26265 and WO 99/03861 to Novo Nordisk A/S which are incorporated herein by reference, insulin sensitizers, DPP-IV (dipeptidyl peptidase-IV) inhibitors, inhibitors of hepatic enzymes involved in stimulation of gluconeogenesis and/or glycogenolysis, glucose uptake modulators, compounds modifying the lipid metabolism such as antihyperlipidemic agents and antilipidemic agents, compounds lowering food intake, PPAR (peroxisome proliferator-activated receptor) and RXR (retinoid X receptor) agonists and agents acting on the ATP-dependent potassium channel of the xcex2-cells.
In one embodiment of the invention the present compounds are administered in combination with insulin.
In a further embodiment the present compounds are administered in combination with a sulphonylurea eg tolbutamide, glibenclamide, glipizide or glicazide.
In another embodiment the present compounds are administered in combination with a biguanide eg metformin.
In yet another embodiment the present compounds are administered in combination with a meglitinide eg repaglinide.
In still another embodiment the present compounds are administered in combination with a thiazolidinedione eg troglitazone, ciglitazone, pioglitazone, rosiglitazone or the compounds disclosed in WO 97/41097 to Dr. Reddy""s Research Foundation.
Furthermore, the present compounds may be administered in combination with the insulin sensitizers disclosed in WO 99/19313 to Dr. Reddy""s Research Foundation.
In a further embodiment the present compounds are administered in combination with an xcex1-glucosidase inhibitor eg miglitol or acarbose.
In another embodiment the present compounds are administered in combination with an agent acting on the ATP-dependent potassium channel of the xcex2-cells eg tolbutamide, glibenclamide, glipizide, glicazide or repaglinide.
Furthermore, the present compounds may be administered in combination with nateglinide.
In still another embodiment the present compounds are administered in combination with an antihyperlipidemic agent or antilipidemic agent eg cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine.
In a further embodiment the present compounds are administered in combination with more than one of the above-mentioned compounds eg in combination with a sulphonylurea and metformin, a sulphonylurea and acarbose, repaglinide and metformin, insulin and a sulphonylurea, insulin and metformin, insulin and troglitazone, insulin and lovastatin, etc.
Furthermore, the compounds according to the invention may be administered in combination with one or more antiobesity agents or appetite regulating agents.
Such agents may be selected from the group consisting of CART (cocaine amphetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melanocortin 4) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releasing factor binding protein) antagonists, urocortin agonists, xcex23 agonists, MSH (melanocyte-stimulating hormone) agonists, MCH (melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin re-uptake inhibitors, serotonin and noradrenaline re-uptake inhibitors, 5HT (serotonin) agonists, bombesin agonists, galanin antagonists, growth hormone, growth hormone releasing compounds, TRH (thyreotropin releasing hormone) agonists, UCP 2 or 3 (uncoupling protein 2 or 3) modulators, leptin agonists, DA (dopamine) agonists (bromocriptin, doprexin), lipase/amylase inhibitors, PPAR modulators, RXR modulators or TR xcex2 agonists.
In one embodiment of the invention the antiobesity agent is leptin.
In another embodiment the antiobesity agent is dexamphetamine or amphetamine.
In another embodiment the antiobesity agent is fenfluramine or dexfenfluramine.
In still another embodiment the antiobesity agent is sibutramine.
In a further embodiment the antiobesity agent is orlistat.
In another embodiment the antiobesity agent is mazindol or phentermine.
Furthermore, the present compounds may be administered in combination with one or more antihypertensive agents. Examples of antihypertensive agents are xcex2-blockers such as alprenolol, atenolol, timolol, pindolol, propranolol and metoprolol, ACE (angiotensin converting enzyme) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril, calcium channel blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and xcex1-blockers such as doxazosin, urapidil, prazosin and terazosin. Further reference can be made to Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, Pa., 1995.
It should be understood that any suitable combination of the compounds according to the invention with one or more of the above-mentioned compounds and optionally one or more pharmacologically active substances are considered to be within the scope of the present invention.
Pharmaceutical Compositions
The compounds of the invention may be administered alone or in combination with pharmaceutically acceptable carriers or excipients, in either single or multiple doses. The pharmaceutical compositions according to the invention may be formulated with pharmaceutically acceptable carriers or diluents as well as any other known adjuvants and excipients in accordance with conventional techniques such as those disclosed in Remington: The Science and Practice of Pharmacy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, Pa., 1995.
The pharmaceutical compositions may be specifically formulated for administration by any suitable route such as the oral, rectal, nasal, pulmonary, topical (including buccal and sublingual), transdermal, intracisternal, intraperitoneal, vaginal and parenteral (including subcutaneous, intramuscular, intrathecal, intravenous and intradermal) route, the oral route being preferred. It will be appreciated that the preferred route will depend on the general condition and age of the subject to be treated, the nature of the condition to be treated and the active ingredient chosen.
Pharmaceutical compositions for oral administration include solid dosage forms such as capsules, tablets, dragees, pills, lozenges, powders and granules. Where appropriate, they can be prepared with coatings such as enteric coatings or they can be formulated so as to provide controlled release of the active ingredient such as sustained or prolonged release according to methods well known in the art.
Liquid dosage forms for oral administration include solutions, emulsions, suspensions, syrups and elixirs.
Pharmaceutical compositions for parenteral administration include sterile aqueous and non-aqueous injectable solutions, dispersions, suspensions or emulsions as well as sterile powders to be reconstituted in sterile injectable solutions or dispersions prior to use. Depot injectable formulations are also contemplated as being within the scope of the present invention.
Other suitable administration forms include suppositories, sprays, ointments, cremes, gels, inhalants, dermal patches, implants etc.
A typical oral dosage is in the range of from about 0.001 to about 100 mg/kg body weight per day, preferably from about 0.01 to about 50 mg/kg body weight per day, and more preferred from about 0.05 to about 10 mg/kg body weight per day administered in one or more dosages such as 1 to 3 dosages. The exact dosage will depend upon the frequency and mode of administration, the sex, age, weight and general condition of the subject treated, the nature and severity of the condition treated and any concomitant diseases to be treated and other factors evident to those skilled in the art.
The formulations may conveniently be presented in unit dosage form by methods known to those skilled in the art. A typical unit dosage form for oral administration one or more times per day such as 1 to 3 times per day may contain of from 0.05 to about 1000 mg, preferably from about 0.1 to about 500 mg, and more preferred from about 0.5 mg to about 200 mg.
For parenteral routes, such as intravenous, intrathecal, intramuscular and similar administration, typically doses are in the order of about half the dose employed for oral administration.
The compounds of this invention are generally utilized as the free substance or as a pharmaceutically acceptable salt thereof. One example is an acid addition salt of a compound having the utility of a free base. When a compound of the formula (I) contains a free base such salts are prepared in a conventional manner by treating a solution or suspension of a free base of the formula (I) with a chemical equivalent of a pharmaceutically acceptable acid, for example, inorganic and organic acids. Representative examples are mentioned above. Physiologically acceptable salts of a compound with a hydroxy group include the anion of said compound in combination with a suitable cation such as sodium or ammonium ion.
For parenteral administration, solutions of the novel compounds of the formula (I) in sterile aqueous solution, aqueous propylene glycol or sesame or peanut oil may be employed. Such aqueous solutions should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. The aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. The sterile aqueous media employed are all readily available by standard techniques known to those skilled in the art.
Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatine, agar, pectin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, polyoxyethylene or water. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. The pharmaceutical compositions formed by combining the novel compounds of the formula (I) and the pharmaceutically acceptable carriers are then readily administered in a variety of dosage forms suitable for the disclosed routes of administration. The formulations may conveniently be presented in unit dosage form by methods known in the art of pharmacy.
Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules or tablets, each containing a predetermined amount of the active ingredient, and which may include a suitable excipient. These formulations may be in the form of powder or granules, as a solution or suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion.
If a solid carrier is used for oral administration, the preparation may be tabletted, placed in a hard gelatine capsule in powder or pellet form or it can be in the form of a troche or lozenge. The amount of solid carrier will vary widely but will usually be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatine capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
A typical tablet that may be prepared by conventional tabletting techniques may contain:
If desired, the pharmaceutical composition of the invention may comprise the compound of the formula (I) in combination with further pharmacologically active substances such as those described in the foregoing.
Experimental
In the following section binding assays as well as functional assays useful for evaluating the efficiency of the compounds of the invention are described.
Binding of compounds to the glucagon receptor was determined in a competition binding assay using the cloned human glucagon receptor.
Antagonism was determined as the ability of the compounds to inhibit the amount of cAMP formed in the presence of 5 nM glucagon.
For further characterization, antagonism was determined in a functional assay, measured as the ability of the compounds to right-shift the glucagon dose-response curve. Using at least 3 different antagonist concentrations, the Ki was calculated from a Schild plot.
Glucagon Binding Assay (I)
Receptor binding was assayed using cloned human receptor (Lok et al., Gene 140, 203-209 (1994)). The receptor inserted in the pLJ6xe2x80x2 expression vector using EcoRI/SSt1 restriction sites (Lok et al.) was expressed in a baby hamster kidney cell line (A3 BHK 570-25). Clones were selected in the presence of 0.5 mg/mL G418 and were shown to be stable for more than 40 passages. The Kd was shown to be 0.1 nM.
Plasma membranes were prepared by growing cells to confluence, detaching them from the surface and resuspending the cells in cold buffer (10 mM tris/HCl, pH 7.4 containing 30 mM NaCl, 1 mM dithiothreitol, 5 mg/L leupeptin (Sigma), 5 mg/L pepstatin (Sigma), 100 mg/L bacitracin (Sigma) and 15 mg/L recombinant aprotinin (Novo Nordisk A/S)), homogenization by two 10-s bursts using a Polytron PT 10-35 homogenizer (Kinematica), and centrifugation upon a layer of 41 w/v % sucrose at 95.000xc3x97g for 75 min. The white band located between the two layers was diluted in buffer and centrifuged at 40.000xc3x97g for 45 min. The precipitate containing the plasma membranes was suspended in buffer and stored at xe2x88x9280xc2x0 C. until use.
Glucagon was iodinated according to the chloramine T method (Hunter and Greenwood, Nature 194, 495 (1962)) and purified using anion exchange chromatography (Jxc3x8rgensen et al., Hormone and Metab. Res. 4, 223-224 (1972). The specific activity was 460 xcexcCi/xcexcg on the day of iodination. Tracer was stored at xe2x88x9218xc2x0 C. in aliquots and were used immediately after thawing.
Binding assays were carried out in triplicate in filter microtiter plates (MADV N65, Millipore). The buffer used in this assay was 50 mM HEPES, 5 mM EGTA, 5 mM MgCl2, 0.005% tween 20, pH 7.4. Glucagon was dissolved in 0.05 M HCl, added an equal amount (w/w) of HSA and freeze-dried. On the day of use, it was dissolved in water and diluted in buffer to the desired concentrations.
Test compounds were dissolved and diluted in DMSO. 140 xcexcL buffer, 25 xcexcL glucagon or buffer, and 10 xcexcL DMSO or test compound were added to each well. Tracer (50.000 cpm) was diluted in buffer and 25 xcexcL were added to each well. 1-4 xcexcg freshly thawed plasma membrane protein diluted in buffer was then added in aliquots of 25 xcexcL to each well. Plates were incubated at 30xc2x0 C. for 2 hours. Non-specific binding was determined with 10xe2x88x926 M of glucagon. Bound tracer and unbound tracer were then separated by vacuum filtration (Millipore vacuum manifold). The plates were washed with 2xc3x97100 xcexcL buffer/well. The plates were air dried for a couple of hours, whereupon the filters were separated from the plates using a Millipore Puncher. The filters were counted in a gamma counter.
Functional Assay (I)
The functional assay was carried out in 96 well microtiter plates (tissue culture plates, Nunc). The resulting buffer concentrations in the assay were 50 mM tris/HCl, 1 mM EGTA, 1.5 mM MgSO4, 1.7 mM ATP, 20 xcexcM GTP, 2 mM IBMX (isobutyl-methyl-xanthine), 0.02% tween-20 and 0.1% HSA. pH was 7.4. Glucagon and proposed antagonist were added in aliquots of 35 xcexcL diluted in 50 mM tris/HCl, 1 mM EGTA, 1.85 mM MgSO4, 0.0222% tween-20 and 0.111% HSA, pH 7.4. 20 xcexcL of 50 mM tris/HCl, 1 mM EGTA, 1.5 mM MgSO4, 11.8 mM ATP, 0.14 mM GTP, 14 mM IBMX and 0.1% HSA, pH 7.4 was added. GTP was dissolved immediately before the assay.
50 xcexcL containing 5 xcexcg of plasma membrane protein was added in a tris/HCl, EGTA, MgSO4, HSA buffer (the actual concentrations were dependent upon the concentration of protein in the stored plasma membranes).
The total assay volume was 140 xcexcL. The assay was incubated for 2 hours at 37xc2x0 C. with continuous shaking. Reaction was terminated by addition of 25 xcexcL 0.5 N HCl. cAMP was measured by the use of a scintillation proximity kit (Amersham).
Glucagon Binding Assay (II)
Receptor binding was assayed using the cloned human receptor (Lok et al., Gene 140, 203-209 (1994)). The receptor inserted in the pLJ6xe2x80x2 expression vector using EcoRI/SSt1 restriction sites (Lok et al.) was expressed in a baby hamster kidney cell line (A3 BHK 570-25). Clones were selected in the presence of 0.5 mg/mL G418 and were shown to be stable for more than 40 passages. The Kd was shown to be 0.1 nM.
Plasma membranes were prepared by growing cells to confluence, detaching them from the surface and resuspending the cells in cold buffer (50 mM tris base, pH 7.4 containing 0.32 mM sucrose, 2 mM EGTA, 1 xcexcg/mL leupeptin, 5 xcexcg/mL pepstatin A, 5 xcexcg/mL aprotinin, 1 mM phenylmethylsulfonylfluoride (all from Sigma)), homogenization by two 10-s bursts using a Polytron PT 10-35 homogenizer (Kinematica), and centrifugation. The homogenate was resuspended and centrifuged again. The final precipitate containing the plasma membranes was suspended in buffer and stored at xe2x88x9280xc2x0 C. until use.
Binding assays were carried out in duplicate in polypropylene tubes or microtiter plates. The buffer used in this assay was 50 mM HEPES pH 7.4 containing 5 mM EGTA, 5 mM MgCl2 and 0.005% Tween 20. Sample (glucagon (Bachem Calif.) or test compounds) was added to each tube or well. Tracer (xcx9c25.000 cpm) was diluted in buffer and was added to each tube or well. 0.5 mg freshly thawed plasma membrane protein diluted in buffer was then added in aliquots to each tube or well. Tubes or plates were incubated at 37xc2x0 C. for 1 hour. Non-specific binding was determined with 10xe2x88x927 M of glucagon. Bound tracer and unbound tracer were then separated by vacuum filtration. The tubes or wells were washed twice with 0.01% Triton X-100 buffer. Scintillation fluid was added to the plates and radioactivity was quantified using a scintillation counter.
Functional Assay (II)
The functional assay determined the ability of the compounds to antagonize glucagon-stimulated formation of cAMP in a whole-cell assay. The assay was carried out in borosilicate glass 12xc3x9775 tubes. The buffer concentrations in the assay were 10 mM HEPES, 1 mM EGTA, 1.4 mM MgCl2, 0.1 mM IBMX, 30 mM NaCl, 4.7 mM KCl, 2.5 mM NaH2PO4, 3 mM glucose and 0.2% BSA. The pH was 7.4. Loose whole cells (0.5 mL, 106/mL) were pretreated with various concentrations of compounds for 10 min at 37xc2x0 C., then challenged with glucagon for 20 min. Some aliquots (500 xcexcL) of cells were treated with test compounds (55 xcexcL) alone to test for agonist activity. The reactions were terminated by centrifugation, followed by cell lysis with the addition of 500 xcexcL 0.1% HCl. Cellular debris was pelleted and the supematant containing cAMP evaporated to dryness. cAMP was measured by the use of an RIA kit (NEN, NEK-033). Some assays were carried out utilizing the adenylate cyclase FlashPlate system from NEN.
Glucagon Binding Assay (III)
BHK (baby hamster kidney cell line) cells were transfected with the human glucagon receptor and a membrane preparation of the cells was prepared. Wheat Germ Agglutinin derivatized SPA beads containing a scintillant (WGA beads) (Amersham) bound the membranes. 125I-glucagon bound to human glucagon receptor in the membranes and excited the scintillant in the WGA beads to light emission. Glucagon or samples binding to the receptor competed with 125I-glucagon.
All steps in the membrane preparation were kept on ice or performed at 4xc2x0 C. BHK cells were harvested and centrifuged. The pellet was resuspended in homogenisation buffer (25 mM HEPES pH=7.4, 2.5 mM CaCl2, 1.0 mM MgCl2, 250 mg/L bacitracin), homogenised 2xc3x9710 sec using Polytron 10-35 homogenizer (Kinematica) and added the same amount of homogenisation buffer as used for resuspension. After centrifugation (15 min at 1000xc3x97g) the supernatant was transferred to cold centrifuge tubes and centrifuged for 45 min at 40.000xc3x97g.
The pellet was resuspended in homogenisation buffer, homogenised 2xc3x9710 sec (Polytron) and additional homogenisation buffer was added. The suspension was centrifuged for 45 min at 40,000xc3x97g and the pellet was resuspended in resuspension buffer (25 mM HEPES pH=7.4, 2.5 mM CaCl2, 1.0 mM MgCl2) and homogenised 2xc3x9710 sec. (Polytron). The protein concentration was normally around 1.75 mg/mL. Stabilisation buffer (25 mM HEPES pH=7.4, 2.5 mM CaCl2, 1.0 mM MgCl2, 1% BSA, 500 mg/L bacitracin, 2.5 M sucrose) was added and the membrane preparation was stored at xe2x88x9280xc2x0 C.
The glucagon binding assay was carried out in opti plates (Polystyrene Microplates, Packard). 50 xcexcL assay buffer (25 mM HEPES pH=7.5, 2.5 mM CaCl2, 1.0 mM MgCl2, 0.003% Tween-20, 0.005% bacitracin, 0.05% sodium azide) and 5 xcexcL glucagon or test compound (in DMSO) were added to each well. 50 xcexcL tracer (125I-porcine glucagon, 70,000 cpm) and 50 xcexcL membranes (12.5 xcexcg) containing the human glucagon receptor were then added to the wells. Finally 50 xcexcL WGA beads containing 1 mg beads were transferred to the well. The assay was incubated for 4 hours on a shaker and then settled for 8-48 hours. The opti plates were counted in a Topcounter. Non-specific binding was determined with 500 nM of glucagon.
Synthesis Methods
The following synthesis protocols refer to intermediate compounds and final products identified in the specification and in the synthetic schemes. The preparation of the compounds of the present invention is described in detail using the following examples, but the chemical reactions described are disclosed in terms of their general applicability to the preparation of the glucagon antagonists of the invention. Occasionally, the reaction may not be applicable as described to each compound included within the disclosed scope of the invention. The compounds for which this occurs will be readily recognised by those skilled in the art. In these cases the reactions can be successfully performed by conventional modifications known to those skilled in the art, that is, by appropriate protection of interfering groups, by changing to other conventional reagents, or by routine modification of reaction conditions. Alternatively, other reactions disclosed herein or otherwise conventional will be applicable to the preparation of the corresponding compounds of the invention. In all preparative methods, all starting materials are known or may easily be prepared from known starting materials. All temperatures are set forth in degrees Celsius and unless otherwise indicated, all parts and percentages are by weight when referring to yields and all parts are by volume when referring to solvents and eluents.
Some of the NMR data shown in the following examples are only selected data.
Unless otherwise specified in the examples, the cis/trans isomeric compounds were obtained as mixtures of cis and trans isomers, which may be separated by chromatography. Thus, the present invention comprises the compounds in form of mixtures of cis and trans isomers as well as the pure isomeric forms.
In the examples the following terms are intended to have the following meanings:
The final products obtained were characterized by analytical RP-HPLC (retention time) and/or by HPLC-MS (molecular mass and/or retention time).
The RP-HPLC analyses were performed on a Waters HPLC system consisting of Waters(trademark) 600S Controller, Waters(trademark) 996 Photodiode Array Detector, Waters(trademark) 717 Autosampler, Waters(trademark) 616 Pump, Waters(trademark) 3 mmxc3x97150 mm 3.5xcexc C-18 Symmetry column and Millenium QuickSet Control Ver. 2.15 using UV detection at 214 nm. A linear gradient was applied from 5% to 90% acetonitrile/0.1% TFA/water over 15 min at a flow rate of 1 mL/minute.
HPLC-MS (Method A)
The following instrumentation was used:
Sciex API 100 Single quadropole mass spectrometer
Perkin Elmer Series 200 Quard pump
Perkin Elmer Series 200 autosampler
Applied Biosystems 785A UV detector
Sedex 55 evaporative light scattering detector
A Valco column switch with a Valco actuator controlled by timed events from the pump.
The Sciex Sample control software running on a Macintosh PowerPC 7200 computer was used for the instrument control and data acquisition.
The HPLC pump was connected to four eluent reservoirs containing:
A: Acetonitrile
B: Water
C: 0.5% TFA in water
D: 0.02 M ammonium acetate
The requirements for samples are that they contain approximately 500 xcexcg/mL of the compound to be analysed in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.)
The analysis was performed at room temperature by injecting 20 xcexcL of the sample solution on the column, which was eluted with a gradient of acetonitrile in either 0.05% TFA or 0.002 M ammonium acetate. Depending on the analysis method varying elution conditions were used.
The eluate from the column was passed through a flow splitting T-connector, which passed approximately 20 xcexcL/min (1/50) through approx. 1 m. 75xcexc fused silica capillary to the API interface of API 100 spectrometer.
The remaining 1.48 mL/min (49/50) was passed through the UV detector and to the ELS detector.
During the LC-analysis the detection data were acquired concurrently from the mass spectrometer, the UV detector and the ELS detector.
The LC conditions, detector settings and mass spectrometer settings used for the different methods are given in the following tables.
HPLC-MS (Method B)
This method was identical to METHOD A but using the following conditions and settings:
HPLC-MS (Method C)
The following instrumentation was used:
Hewlett Packard series 1100 MSD G1946A Single quadropole mass spectrometer
Hewlett Packard series 1100 MSD G1312A Bin pump
Hewlett Packard series 1100 MSD G1313A ALS autosampler
Hewlett Packard series 1100 MSD G1315A DAD diode array detector
The HP LC/MSD ChemStation control software running on a HP Vectra computer was used for the instrument control and data acquisition.
The HPLC pump was connected to two eluent reservoirs containing:
A: 0.05% TFA in water
B: Acetonitrile
The analysis was performed at room temperature by injecting 1 xcexcL of the sample solution on the column which was eluted with a gradient of acetonitrile in 0.05% TFA.
The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
HPLC-MS (Method D)
The following instrumentation was used:
Hewlett Packard series 1100 G1312A Bin Pump
Hewlett Packard series 1100 G1315A DAD diode array detector
Sciex 300 triplequadropole mass spectrometer
Gilson 215 micro injector
Sedex 55 evaporative light scattering detector
Pumps and detectors were controlled by MassChrom 1.1.1 software running on a Macintosh G3 computer. Gilson Unipoint Version 1.90 controls the auto-injector.
The HPLC pump was connected to two eluent reservoirs containing:
A: 0.01% TFA in water
B: 0.01% TFA in acetonitrile
The analysis was performed at room temperature by injecting an appropriate volume of the sample (preferably 1 xcexcL) onto the column that was eluted with a gradient of acetonitrile.
The HPLC conditions, detector settings and mass spectrometer settings used are:
HPLC-MS (Method E)
The following instrumentation was used:
Hewlett Packard series 1100 MSD G1946A Single quadropole mass spectrometer
Hewlett Packard series 1100 MSD G1312A Bin pump
Hewlett Packard series 1100 MSD G1313A ALS autosampler
Hewlett Packard series 1100 MSD G1315A DAD diode array detector
The HP LC/MSD ChemStation control software running on a HP Vectra computer was udes for the instrument control and data acquisition.
The HPLC pump was connected to two eluent reservoirs containing:
A: 0.01% TFA in water
B: Acetonitrile
The analysis was performed at room temperature by injecting 1 xcexcof the sample solution on the column which was eluted with a gradient of acetonitrile in 0.01% TFA.
The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
HPLC-MS (Method F)
The following instrumentation was used:
Hewlett Packard series 1100 G1312A Bin Pump
Hewlett Packard series 1100 Column compartment
Hewlett Packard series 1100 G1315A DAD diode array detector
Hewlett Packard series 1100 MSD
The instrument was controlled by HP Chemstation software.
The HPLC pump was connected to two eluent reservoirs containing:
A: 0.01% TFA in water
B: 0.01% TFA in acetonitrile
The analysis was performed at 40xc2x0 C. by injecting an appropriate volume of the sample (preferably 1 xcexcL) onto the column, which was eluted with a gradient of acetonitrile.
The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
The compounds of the present invention may be purified using one of the following HPLC methods:
Hit Fractionation Method I
HPLC purification/fractionation of hits is performed on Nucleosil C-18 7 xcexcm 8xc3x97100 mm columns (packed by Grom). A standard gradient with water/acetonitrile added 0.01% TFA is used. Flow rate 9 mL/min starting at 10% organic modifier ending after 18 min on 100% organic modifier. This condition is kept for 1 min. Fractions of 4 mL are collected in a deep well collection plate.
Equipment
2 Gilson 306 pumps equipped with 25 mL SC pump heads, Gilson 806 manometer and Gilson 811c dynamic mixing chamber. UV detection is performed with Gilson 119 UV/VIS detector. Gilson 215 Nebula is used as combined injector and fraction collector.
Hit Fractionation Method II
HPLC purification/fractionation of hits is performed on Waters Xterra columns MS C18 5 xcexcm 7.8xc3x97100 mm columns. A standard gradient with water/acetonitrile added 0.01% TFA is used. Flow rate 15 mL/min starting at 10% organic modifier ending after 11 min on 100% modifier. This condition is kept for 1 min. Fractions of 4 mL are collected in a deep well collection plate.
Equipment
1 Gilson 321 pump equipped with 15 mL H1 pumping heads. UV detection is performed with Gilson 119 UV/VIS detector. Gilson 215 Nebula is used as combined injector and fraction collector.
General Procedure (A) for the Solid Phase Synthesis of Compounds of the General Formula (Ia) 
wherein
R1, Z, E and D are as defined for formula (I),
A is 
xe2x80x83wherein
R8 and R9 are as defined for formula (I),
X is 
xe2x80x83wherein
r is as defined for formula (I),
Lea is a leaving group such as chloro, bromo, iodo, mesyl or tosyl,
Leaxe2x80x2 is a leaving group such as xe2x80x94OSu, chloro, phenoxy or 4-nitrophenoxy, and
Resin denotes a polystyrene resin with a linker such as the Wang linker: 
wherein PS denotes polystyrene.
Step A
The reaction is known (Wang S. J., J. Am. Chem. Soc. 95, 1328, 1973) and is generally performed by stirring polystyrene resin loaded with a linker such as the Wang linker with a 4-10 molar excess of Fmoc-protected amino acid activated with a 2-5 molar excess of diisopropylcarbodiimide or dicyclohexylcarbodiimide in the presence of a catalyst such as N,N-4-dimethylaminopyridine. The esterification is carried out in a solvent such as THF, dioxane, toluene, dichloromethane, DMF, NMP or a mixture of two or more of these. The reactions are performed between 0xc2x0 C. to 80xc2x0 C., preferably between 20xc2x0 C. to 40xc2x0 C. When the esterification is complete excess of reagents is removed by filtration. The resin is successively washed with the solvent used in the reaction, followed by washings with methanol. The resin bound product can be further dried and analyzed.
Step B
N-Fluorenylmethyloxycarbonyl protection group is removed by treating the resin bound derivative with a 20%-50% solution of a secondary amine such as piperidine in a polar solvent such as DMF or NMP (Carpino L., Han G., J. Org. Chem. 37, 3404, 1972). The reaction is performed between 20xc2x0 C. and 180xc2x0 C., preferably between 20xc2x0 C. and 40xc2x0 C. The deprotection can be quantitated by the absorbance of piperidine-dibenzofulvene adduct released from the resin (The combinatorial index, Ed. Bunin B. A., 1998, Academic press, p. 219). When the reaction is complete excess of reagents is removed by filtration. The resin is successively washed with solvent used in the reaction. The resulting resin bound intermediate is acylated with acid (II). The acylation is known (The combinatorial index, Ed. Bunin B. A., 1998, Academic press, p. 78) and is generally performed by stirring resin bound intermediate with a 2-5 molar excess of acid (II) activated with a 2-5 molar excess of diisopropylcarbodiimide or dicyclohexylcarbodiimide in the presence of a side reaction inhibitor such as N-hydroxybenzotriazole. The acylation is carried out in a solvent such as THF, dioxane, toluene, dichloromethane, DMF, NMP or a mixture of two or more of these. The reactions are performed between 0xc2x0 C. and 80xc2x0 C., preferably between 20xc2x0 C. and 40xc2x0 C. When the acylation is complete excess of reagents is removed by filtration. The resin is successively washed with the solvent used in the reaction, followed by washings with methanol. The resin bound product can be further dried and analyzed.
Step C
The reaction is known (The combinatorial index, Ed. Bunin B. A., 1998, Academic press, p. 112) and is generally performed by stirring the resin bound intermediate obtained in step B with a 10-20 molar excess of amine. The nucleophilic displacement is carried out in a solvent such as DMSO, DMF, NMP or a mixture of two or more of these. The reaction is performed between 20xc2x0 C. and 120xc2x0 C., preferably between 60xc2x0 C. and 80xc2x0 C. When the reaction is complete excess of reagents is removed by filtration. The resin is successively washed with the solvent used in the reaction, followed by washings with methanol. The resin bound product can be further dried and analyzed.
Step D
The reaction is known (The combinatorial index, Ed. Bunin B. A., 1998, Academic press, p. 78) and is generally performed by stirring the resin bound intermediate obtained in step C with a 4-10 molar excess of acid HO-X-D activated with a 2-5 molar excess of diisopropylcarbodiimide or dicyclohexylcarbodiimide in the presence of a catalyst such as pyridine and/or 4-dimethylaminopyridine. The reaction is carried out in a solvent such as THF, dioxane, toluene, dichloromethane, DMF, NMP or a mixture of two or more of these. The reactions are performed between 0xc2x0 C. and 80xc2x0 C., preferably between 20xc2x0 C. and 40xc2x0 C. When the reaction is complete excess of reagents is removed by filtration. Alternatively, a solution of Leaxe2x80x2-X-D in an appropriate solvent such as acetonitrile, toluene, DMF, NMP, THF, dichloromethane, 1,2-dichloroethane or DMSO or a mixture of two or more of these, is added and the mixture is vortexed in the presence of a base such as triethylamine, diisopropylethylamine, dicyclohexylmethylamine or any other tertiary amine or potassium carbonate under heating, if necessary. The resin is successively washed with solvent used in the reaction, followed by washings with dichloromethane.
Step E
The reaction is known (The combinatorial index, Ed. Bunin B. A., 1998, Academic press, p. 21) and is generally performed by stirring resin bound intermediate obtained in step D with a 50-95% solution of TFA. The final cleavage is carried out in a solvent such as THF, dichloromethane, 1,2-dichloroethane, 1,3-dichloropropane, toluene or a mixture of two or more of these. The reaction is performed between 0xc2x0 C. and 80xc2x0 C., preferably between 20xc2x0 C. and 40xc2x0 C. When the reaction is complete the product is removed by filtration. The resin is successively washed with dichloromethane. The product and washings are collected. The solvent is removed and the product is dried in vacuo. The residue is dissolved in a 1:1 mixture of methanol and dichloromethane (1 mL) and concentrated in vacuo. The product (Ia) is dried in vacuo overnight.
General Procedure (B) for the Solid Phase Synthesis of Compounds of the General Formula (Ia):
Alternatively, steps B and C of procedure (A) can be modified so that step C is a reductive amination of a resin bound aldehyde or ketone: 
wherein
R1, Z, E and D are as defined for formula (I),
A is 
xe2x80x83wherein
R8 and R9 are as defined for formula (I),
X is 
xe2x80x83wherein
r is as defined for formula (I),
Leaxe2x80x2 is a leaving group such as xe2x80x94OSu, chloro, phenoxy or 4-nitrophenoxy, and
Resin denotes a polystyrene resin with a linker such as the Wang linker: 
wherein PS denotes polystyrene.
Step B
This step is identical to step B of general procedure (A) with the modification that acid (III) is used instead of acid (II).
Step C
The reaction is generally known (The combinatorial index, Ed. Bunin, B. A. 1998, Academic Press, p. 133) and is generally performed by stirring resin bound aldehyde or ketone with an excess of amine at low pH (by addition of an acid, such as acetic acid or formic acid) in a solvent such as THF, DMF, NMP, methanol, ethanol, DMSO, dichloromethane, 1,2-dichloroethane, trimethyl orthoformate, triethyl orthoformate, or a mixture of two or more of these. A reducing agent such as sodium cyanoborohydride may be used. The reaction is performed between 20xc2x0 C. and 120xc2x0 C., preferably at 25xc2x0 C.